The National Institute of Diabetes & Digestive & Kidney Diseases

Our laboratory is interested in membrane and protein recycling within eukaryotic cells. The approach we are taking is to examine the structure and function of proteins involved in these processes by transmission electron microscopy, including cyro-electron microscopy, and by biochemical methods. Presently, we are examining a 100 kDa GTPase called dynamin, which is involved in the constriction and fission of clathrin-coated pits from plasma membrane during receptor-mediated endocytosis and during membrane retrieval in nerve terminals. We have shown that dynamin self-assembles into spirals and onto lipid bilayers forming dynamin decorated lipid tubes that undergo a large conformational change upon GTP addition resulting in membrane constriction. To further understand the mechanism of dynamin-induced membrane constriction, we have solved the 3-dimensional structure of dynamin in the non-constricted and constricted states by high-resolution electron microscopy and image processing methods. This work suggests a GTP-induced conformational change within the dynamin oligomer is responsible for constriction. Additional dynamin family members have been implicated in numerous fundamental cellular processes, including other membrane fission events, anti-viral activity, cell plate formation and chloroplast biogenesis. We are currently examining the effects of GTP on these proteins to determine if a common mechanism of action exists for all dynamin family members.

Chappie JS, Mears JA, Fang S, Leonard M, Schmid SL, Milligan RA, Hinshaw JE, Dyda F. A pseudoatomic model of the dynamin polymer identifies a hydrolysis-dependent powerstroke.Cell (147): 209-22. 2011. [Full Text/Abstract]

Sundborger A, Soderblom C, Vorontsova O, Evergren E, Hinshaw JE, Shupliakov O. An endophilin-dynamin complex promotes budding of clathrin-coated vesicles during synaptic vesicle recycling.J Cell Sci (124): 133-43, 2011. [Full Text/Abstract]

Mears JA, Lackner LL, Fang S, Ingerman E, Nunnari J, Hinshaw JE. Conformational changes in Dnm1 support a contractile mechanism for mitochondrial fission. Nat Struct Mol Biol (18): 20-6, 2011. [Full Text/Abstract]

Kitajiri S, Sakamoto T, Belyantseva IA, Goodyear RJ, Stepanyan R, Fujiwara I, Bird JE, Riazuddin S, Riazuddin S, Ahmed ZM, Hinshaw JE, Sellers J, Bartles JR, Hammer JA 3rd, Richardson GP, Griffith AJ, Frolenkov GI, Friedman TB. Actin-bundling protein TRIOBP forms resilient rootlets of hair cell stereocilia essential for hearing. Cell (141): 786-98, 2010. [Full Text/Abstract]

Ban T, Heymann JA, Song Z, Hinshaw JE, Chan DC. OPA1 disease alleles causing dominant optic atrophy have defects in cardiolipin-stimulated GTP hydrolysis and membrane tubulation. Hum Mol Genet (19): 2113-22, 2010. [Full Text/Abstract]

Schulz TA, Choi MG, Raychaudhuri S, Mears JA, Ghirlando R, Hinshaw JE, Prinz WA. Lipid-regulated sterol transfer between closely apposed membranes by oxysterol-binding protein homologues. J Cell Bio (187): 889-903, 2009. [Full Text/Abstract]

Heymann JA, Hinshaw JE. Dynamins at a glance. J Cell Sci (122): 3427-31, 2009. [Full Text/Abstract]

Bethoney KA, King MC, Hinshaw JE, Ostap EM, Lemmon MA. A possible effector role for the pleckstrin homology (PH) domain of dynamin. Proc Natl Acad Sci USA (106): 13359-64, 2009. [Full Text/Abstract]

Mears, J. A. & Hinshaw, J. E. Visualization of dynamins. Methods Cell Biol 88, 237-256 (2008). [Full Text/Abstract]

Cassidy-Stone, A. et al. Chemical inhibition of the mitochondrial division dynamin reveals its role in bax/bak-dependent mitochondrial outer membrane permeabilization. Dev Cell 14, 193-204 (2008).[Full Text/Abstract]

Mears, J. A., Ray, P. & Hinshaw, J. E. A corkscrew model for dynamin constriction. Structure 15, 1190-1202 (2007).[Full Text/Abstract]

Nie Z, Hirsch DS, Luo R, Jian X, Stauffer S, Cremesti A, Andrade J, Lebowitz J, Marino M, Ahvazi B, Hinshaw JE, Randazzo PA A BAR domain in the N terminus of the Arf GAP ASAP1 affects membrane structure and trafficking of epidermal growth factor receptor. Curr Biol (16): 130-9, 2006. [Full Text/Abstract]

Naylor K, Ingerman E, Okreglak V, Marino M, Hinshaw JE, Nunnari J Mdv1 interacts with assembled dnm1 to promote mitochondrial division. J Biol Chem (281): 2177-83, 2006. [Full Text/Abstract]

Kochs G, Reichelt M, Danino D, Hinshaw JE, Haller O Assay and functional analysis of dynamin-like Mx proteins. Methods Enzymol (404): 632-43, 2005. [Full Text/Abstract]

Ingerman E, Perkins EM, Marino M, Mears JA, McCaffery JM, Hinshaw JE, Nunnari J Dnm1 forms spirals that are structurally tailored to fit mitochondria. J Cell Biol (170): 1021-7, 2005. [Full Text/Abstract]

Hickman AB, Perez ZN, Zhou L, Musingarimi P, Ghirlando R, Hinshaw JE, Craig NL, Dyda F Molecular architecture of a eukaryotic DNA transposase. Nat Struct Mol Biol (12): 715-21, 2005. [Full Text/Abstract]

Danino D, Moon KH, Hinshaw JE Rapid constriction of lipid bilayers by the mechanochemical enzyme dynamin. J Struct Biol (147): 259-67, 2004. [Full Text/Abstract]

Chen YJ Zhang P Egelman EH Hinshaw JE The stalk region of dynamin drives the constriction of dynamin tubes. Nat Struct Mol Biol (11): 574-5, 2004. [Full Text/Abstract]

Hinshaw JE Milligan RA Nuclear pore complexes exceeding eightfold rotational symmetry. J Struct Biol (141): 259-68, 2003. [Full Text/Abstract]

Danino D Hinshaw JE Dynamin family of mechanoenzymes. Curr Opin Cell Biol (13): 454-60, 2001. [Full Text/Abstract]

Zhang P Hinshaw JE Three-dimensional reconstruction of dynamin in the constricted state. Nat Cell Biol (3): 922-6, 2001. [Full Text/Abstract]

Hinshaw JE Dynamin and its role in membrane fission. Annu Rev Cell Dev Biol (16): 483-519, 2000. [Full Text/Abstract]

Hinshaw JE Dynamin spirals. Curr Opin Struct Biol (9): 260-7, 1999. [Full Text/Abstract]

Sweitzer SM Hinshaw JE Dynamin undergoes a GTP-dependent conformational change causing vesiculation. Cell (93): 1021-9, 1998. [Full Text/Abstract]

Carr JF Hinshaw JE Dynamin assembles into spirals under physiological salt conditions upon the addition of GDP and gamma-phosphate analogues. J Biol Chem (272): 28030-5, 1997. [Full Text/Abstract]

Warnock DE Hinshaw JE Schmid SL Dynamin self-assembly stimulates its GTPase activity. J Biol Chem (271): 22310-4, 1996. [Full Text/Abstract]

Baba T, Damke H, Hinshaw JE, Ikeda K, Schmid SL, Warnock DE Role of dynamin in clathrin-coated vesicle formation. Cold Spring Harb Symp Quant Biol (60): 235-42, 1995. [Full Text/Abstract]

Hinshaw JE Architecture of the nuclear pore complex and its involvement in nucleocytoplasmic transport. Biochem Pharmacol (47): 15-20, 1994. [Full Text/Abstract]

Hinshaw JE Carragher BO Milligan RA Architecture and design of the nuclear pore complex. Cell (69): 1133-41, 1992. [Full Text/Abstract]